TLR-3 expression and isotype controls are represented by the white and the grey layers, respectively. Click here to view.(233K, pptx) Fig. (NF)-B and AP-1 in peripheral blood mononuclear cells (PBMC). Activation of (a) NF-B and (b) AP-1 was measured by enzyme-linked immunosorbent assay (ELISA) in nuclear extracts of PBMC from 14 controls, 13 patients in relapse, 13 patients in remission and 13 patients in remission after rituximab therapy (remission RTX+). No significant difference was found between the four groups. cei0182-0332-sd2.pptx (51K) GUID:?E47E3402-2DCD-4329-B171-2E78A1AEDCAB Table S1. Blood cell count of patients with idiopathic nephrotic syndrome and controls. Table S2. Quantity of cells analysed by circulation cytometry in isolated peripheral blood mononuclear cells (PBMC) subsets of patients with idiopathic nephrotic syndrome and controls. cei0182-0332-sd3.docx (15K) GUID:?2DF4070D-F137-42F9-A556-C58E10F56A9A Abstract The efficacy of steroids and immunosuppressive treatments in idiopathic nephrotic syndrome (INS) hints at the implication of immune cells in the pathophysiology of the disease. Toll-like receptor (TLR) dysfunctions are involved in many kidney diseases of immune origin, but remain little explained in INS. We investigated the expression and function of TLRs in peripheral blood mononuclear cells (PBMC) of INS children, including 28 in relapse, 23 in remission and 40 controls. No child experienced any sign of contamination, but a Valrubicin higher EpsteinCBarr computer virus viral weight was measured in the PBMC of relapsing patients. TLR-3 expression was increased in B cells only during INS remission. There was a negative correlation between proteinuria and TLR-3 expression in total and the main subsets of PBMC from INS patients. The expression of TLR-8 was also increased in both CD4+ T cells and B cells in INS remission. There was a negative correlation between proteinuria and TLR-8 expression in total PBMC, CD4+ T cells and B Valrubicin cells of INS patients. Nevertheless, TLR-3 and TLR-8 expression was normalized in all PBMC subsets in an additional group of 15 INS patients in remission with B cell repletion after rituximab therapy. Paradoxically, interferon (IFN) regulatory factor 3 transactivation was increased in PBMC of all INS patients. secretion of IFN- and interleukin 6 were increased spontaneously in PBMC of INS remission patients, whereas PBMC from all INS patients displayed an impaired IFN- secretion after TLR-3 activation. Thus, TLR-3 pathway dysfunctions may be closely involved in INS pathogenesis. Keywords: blood leucocytes, cytokines, immunoglobulins, proteinuria, steroid-sensitive nephrotic syndrome Introduction Idiopathic nephrotic syndrome (INS) accounts for 90% of all glomerular nephropathies in child years, and is usually characterized by massive proteinuria and Valrubicin hypoalbuminaemia 1. The disease affects the kidney exclusively and is marked typically by the effacement of podocyte foot processes without glomerular deposit or inflammatory lesion. The clinical course of INS is largely dependent upon the response to steroids. Relapses are frequent after withdrawal of steroid treatment, with up to 60% of patients being steroid-dependent. Immunosuppressive treatments, such as cyclophosphamide and calcineurin inhibitors, are administered as a second-line treatment for INS to prevent relapses and to spare steroid usage 2. Valrubicin Rituximab (RTX), a chimeric humanCmouse anti-CD20 monoclonal antibody, is also effective in the treatment of INS in preventing relapses in steroid-dependent patients 3,4. The efficacy of immunosuppressive treatments indicates that INS may be associated with dysfunctions of the immune system 5. The main subsets of immune cells have all been linked to the pathogenesis of INS. The injection of CD34+ Rabbit polyclonal to AGBL2 peripheral blood mononuclear cells (PBMC), but not CD34? PBMC, from relapsing INS patients induces albuminuria and podocyte foot effacement in immunodeficient mice 6. Immature CD34+ circulating cells are expanded during INS relapses 7, as well as several T cell populations such as memory T cells 8 and interleukin (IL)-17-generating CD4+ T cells, associated with a reduced number and activity of regulatory T cells 9,10. Numerous studies have concluded that INS patients present an imbalance between T helper type 1 (Th1) and Th2 responses, with a pattern towards a.